Tapio Ilma, Fischer Daniel, Blasco Lucia, Tapio Miika, Wallace R John, Bayat Ali R, Ventto Laura, Kahala Minna, Negussie Enyew, Shingfield Kevin J, Vilkki Johanna
Green Technology, Natural Resources Institute Finland, Jokioinen, Finland.
Bio-based business and industry, Natural Resources Institute Finland, Jokioinen, Finland.
PLoS One. 2017 Jul 13;12(7):e0180260. doi: 10.1371/journal.pone.0180260. eCollection 2017.
The ruminal microbiome, comprising large numbers of bacteria, ciliate protozoa, archaea and fungi, responds to diet and dietary additives in a complex way. The aim of this study was to investigate the benefits of increasing the depth of the community analysis in describing and explaining responses to dietary changes. Quantitative PCR, ssu rRNA amplicon based taxa composition, diversity and co-occurrence network analyses were applied to ruminal digesta samples obtained from four multiparous Nordic Red dairy cows fitted with rumen cannulae. The cows received diets with forage:concentrate ratio either 35:65 (diet H) or 65:35 (L), supplemented or not with sunflower oil (SO) (0 or 50 g/kg diet dry matter), supplied in a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments and four 35-day periods. Digesta samples were collected on days 22 and 24 and combined. QPCR provided a broad picture in which a large fall in the abundance of fungi was seen with SO in the H but not the L diet. Amplicon sequencing showed higher community diversity indices in L as compared to H diets and revealed diet specific taxa abundance changes, highlighting large differences in protozoal and fungal composition. Methanobrevibacter ruminantium and Mbb. gottschalkii dominated archaeal communities, and their abundance correlated negatively with each other. Co-occurrence network analysis provided evidence that no microbial domain played a more central role in network formation, that some minor-abundance taxa were at nodes of highest centrality, and that microbial interactions were diet specific. Networks added new dimensions to our understanding of the diet effect on rumen microbial community interactions.
瘤胃微生物群由大量细菌、纤毛虫原生动物、古菌和真菌组成,对饮食和膳食添加剂的反应方式复杂。本研究的目的是探讨在描述和解释对饮食变化的反应时增加群落分析深度的益处。对从四头装有瘤胃瘘管的多胎北欧红牛获得的瘤胃消化物样本进行了定量PCR、基于小亚基核糖体RNA(ssu rRNA)扩增子的分类群组成、多样性和共现网络分析。奶牛接受的日粮中粗饲料与精饲料的比例分别为35:65(高比例日粮,即H日粮)或65:35(低比例日粮,即L日粮),并分别添加或不添加葵花籽油(SO)(0或50克/千克日粮干物质),采用4×4拉丁方设计,处理因素为2×2析因排列,共四个35天周期。在第22天和第24天收集消化物样本并合并。定量PCR提供了一个大致情况,即在H日粮中添加SO时真菌丰度大幅下降,而在L日粮中未出现这种情况。扩增子测序显示,与H日粮相比,L日粮的群落多样性指数更高,并揭示了日粮特异性分类群丰度变化,突出了原生动物和真菌组成的巨大差异。瘤胃甲烷短杆菌和戈氏甲烷短杆菌在古菌群落中占主导地位,它们的丰度彼此呈负相关。共现网络分析提供的证据表明,没有一个微生物域在网络形成中发挥更核心的作用,一些低丰度分类群处于最高中心度的节点,并且微生物相互作用具有日粮特异性。网络为我们理解日粮对瘤胃微生物群落相互作用的影响增加了新的维度。